title: Phanta: a non-fluorescent photochromic acceptor for pcFRET.
abstract: We have developed an orange non-fluorescent photochromic protein (quantum yield, 0.003) we call Phanta that is useful as an acceptor in pcFRET applications. Phanta can be repeatedly inter-converted between the two absorbing states by alternate exposure to cyan and violet light. The absorption spectra of Phanta in one absorbing state shows excellent overlap with the emission spectra of a number of donor green fluorescent proteins including the commonly used EGFP. We show that the Phanta-EGFP FRET pair is suitable for monitoring the activation of caspase 3 in live cells using readily available instrumentation and a simple protocol that requires the acquisition of two donor emission images corresponding to Phanta in each of its photoswitched states. This the first report of a genetically encoded non-fluorescent acceptor for pcFRET.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton Campus, Melbourne, Victoria, Australia.
authors.forename: Craig
authors.initials: C
authors.lastname: Don Paul
authors.forename: Csaba
authors.initials: C
authors.lastname: Kiss
authors.forename: Daouda A K
authors.initials: DA
authors.lastname: Traore
authors.forename: Lan
authors.initials: L
authors.lastname: Gong
authors.forename: Matthew C J
authors.initials: MC
authors.lastname: Wilce
authors.forename: Rodney J
authors.initials: RJ
authors.lastname: Devenish
authors.forename: Andrew
authors.initials: A
authors.lastname: Bradbury
authors.forename: Mark
authors.initials: M
authors.lastname: Prescott
mesh_terms: D000595:Amino Acid Sequence; D015374:Biosensing Techniques / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010777:Photochemistry / Q000379:methods*; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins